Effect of diets containing fish protein hydrolisates on growth and immune performance of asian seabass

EFFECT OF DIETS CONTAINING FISH PROTEIN HYDROLISATES ON GROWTH AND
IMMUNE PERFORMANCE OF ASIAN SEABASS (Lates calcarifer Bloch) WHEN
REARED IN FARM CONDITIONS
Romi Novriadi1
, Tinggal Hermawan1
, Muh Kadari1
, Dikrurrahman1
, Ibtisam2,
Mikael
Herault3
, Vincent Fournier3
dan Paul Seguin3
1)
Batam Mariculture Development Center, Directorate General of Aquaculture, Ministry of Marine Affairs
and Fisheries, Republic of Indonesia
2)
Master Student at Human Nutrion and Rural Development, Faculty of Bioscience Engineering,
University of Ghent
3)
Aquativ (DIANA Division, Member of SYMRISE Group), ZA du Gohélis 56250 Elven, France
*: Corresponding author : Romi Novriadi, Jl. Raya Barelang Jembatan III, Pulau Setoko, Batam – 29422.
E-mail: Romi_bbl@yahoo.co.id
A B S T R A C T
We investigated the effects of fish protein hydrolysate (FPH) on zootechnical performance and
immune response of the Asian Seabass Lates calcarifer Bloch. Experimental fish were fed with
3 diets: a local commercial diet (control), coated or not, with 2 and 3% FPH (w/w). Twelve
thousand Asian Seabass juveniles (5.88±0.56 g) were divided into three groups and two
replicates reared in nursery tanks (2000 L). The remaining fish were then used for grow-out
experiment in floating net cages (1m x 1 m x 3 m). Zootechnical performances were assessed
at both stages with following indicators: total weight gain (TWG), % relative weight gain (%
RWG), % specific growth rate (% SGR), final weight (g) and final length (cm). At the end of each
trial period, fish immune status was assessed through blood sampling and the measurement of
Neutrophile (%), Monocyte (%), Lymphocyte (%), Macrophage (105
cell/mL), Leukocyte (103
cell/mL) and Phagocytes activity (%). At the end of the nursery trial, an immersion bacterial
challenge with Vibrio parahaemolyticus (105
cells mL-1
) was implemented. The results showed
that dietary FPH supplementation significantly influenced the growth and immune status of
Asian Seabass when compared to the control group. Fish fed FPH supplemented diet yielded
higher growth rates and survival rates than non supplemented group. Fish phagocytic activity
and resistance to a bacterial challenge were also improved by dietary FPH supplementation.
These results may be related to the significant changes observed in fish leukocyte profiles,
when fed FPH supplemented diets. Altogether, these results show the positive contribution of
FPH to the sustainability of Asian seabass farming.
Keywords: Functional hydrolysates, Asian Seabass, Zootechnical performance, Immune
response, Phagocytic activity, Nursery, Grow out
INTRODUCTION
In Asian Seabass culture, disease
outbreaks are being increasingly reported as
a major constraint to the sustainable growth
of production (Chong et al., 1987; Bloch and
Larsen, 1993; Chou et al., 1998). Many
diseases are linked to the stress conditions
associated with the intensification systems
of farming and the degradation of the
environmental quality. Under poor
conditions, live food and larvi are often
opportunistically infected by fungi, bacteria
and viruses. Conventional treatment of
aquatic diseases through the use of
disinfectants and antibiotics to overcome the
bacterial infection problem are having limited
value and has stimulated the development
of bacterial resistance (Defoirdt et al., 2007;
Subasinghe, 1997; Cabello, 2006). Another
problem created by the unrestricted use of
antibiotics is the presence of residual
antibiotics in commercialized aquaculture
products (Saitanu et al., 1994; Grave et al.,
1996; 1999; Goldburg et al., 2001; European
Commission, 2001a,b). This problem has
led to allergy and toxicity in humans
(Alderman and Hastings, 1998; Cabello,
2006) and resulted in shifts in the diversity of
the microbiota due to the permanent
existence of large amounts of antibiotics in
the environment (Cabello, 2003).
Consequently, novel preventive approaches
in aquaculture are urgently needed, e.g.
vaccines, immunostimulants and probiotics
(Marques et al., 2006). Currently, the
industry goes towards much more holistic
approach consisting in protecting aquatic
animals from diseases without the use of
antibiotics by enhancing the resistance of

cultured fish to diseases (Defoirdt et al.,
2007; Smith, V.J et al., 2003) and the use of
enzymatic hydrolysates processed from fish
by-products and showing immunostimulating
properties is one of this promising holistic
approach (Cook et al., 2003).
Over recent years, fish protein
hydrolysates (FPH) have been used as
ingredients for marine fish diets (Ouellet et
al., 1997; Aguila et al., 2007), as a naturally
high source of digestible nutrients, low
molecular weight compounds such as
nucleotides, amino acid and derivatives and
bioactive peptides showing antioxidative,
hormone like, antistress or antimicrobial
activities (Klompong et al., 2007;
Thiansilakul et al., 2007; Liaset and Espe,
2008). Several studies have investigated the
effects of FPH on growth performance and
nonspecific immunity of several fish species
(Berge and Storebakken, 1996; Bøgwald et
al., 1996; Carvalho et al., 1997; Liang et al.,
2006). However, to our knowledge, no
similar studies have been done on the Asian
Seabass Lates calcarifer, especially at two
different rearing periods, namely: the
nursery phase and grow-out phase.
Therefore, the aim of the present study was
to evaluate the effects of dietary
supplementation with functional
hydrolysates on zootechnical and immune
performance in L. calcarifer when reared in
farm conditions.
MATERIALS AND METHODS
Fish protein hydrolysates (FPH)
Fish protein hydrolysate (FPH)
obtained from fishery co-products
(AQUATIV, SPF Diana, Batch No.
203140305, ACTIPAL HL1, krill and tuna
hydrolysate liquid for aquafeed) was used in
the experiments and stored at 40
C until the
end of experiment.
Experimental Animals
1-2 g of L. calcarifer) obtained from
Batam Mariculture Development Center
were used as the test animals. The culture
density at the nursery phase was 2000
fish/m3
and 500 fish at 1 x 1 x 3 m floating
net cages. Acclimatization process was
conducted for 2 weeks preceding the
experiment and 80 – 100 % of water
renewal was performed daily.
Experimental Design and Diet
Administration
Both nursery and grow-out trials were
implemented at the Batam Mariculture
Development Center for 6 weeks and 9
weeks respectively. Experimental design
consisted in graded levels of supplemented
hydrolysate (0, 2 and 3% w/w) coated on a
local extruded commercial diet using a
cement mixer. Each experimental diet was
randomly allocated to 2 replicate tanks
(2000 L capacity) or cages (1 x 1 x 3 m) for
nursery and grow-out rearing respectively.
Feeding rates were fixed to 7% of the
biomass equally distributed in 3 meals over
12 hours, with daily adjustments based on
mortality rates and growth model
assumptions. The experimental tanks were
provided with continuous aeration and water
was changed daily before feeding. Tanks
were cleaned and uneaten feed was
collected 2h after the feeding. Water quality
parameters such as temperature, pH,
salinity and dissolved oxygen were daily
monitored using portable instruments, while
critical parameters such as total ammonia
(NH3) and nitrite (NO2) were measured on
alternate days following standard methods.
The trials were terminated when fish
reached 20 g and 50 to 60 g during the
nursery and grow-out periods respectively.
Zootechnical Performances
Growth performance was expressed
as the total weight gain (TWG), relative
weight gain (RWG) and specific growth rate
(SGR). The calculation formulas were as
follows:
TWG (g) = Wt − Wi,
RWG (%) = ( Wt – Wi ) × 100 / Wi,
SGR (%) = ( lnWt – lnWi )× 100 / d,
Survival (%) = ( number of fish harvested /
number of fish stocked ) × 100.
Feed conversion ratio (FCR) = total amount
of feed consumed (kg) / biomass increase
(kg).
where Wi and Wt are the initial and final
mean weights (g), respectively, and d
represents the number of feeding days
(Mojjada et al., 2013).
Blood sampling and analysis
At the end of the trial, ten fish per
group (five fish randomly captured from
each cage) were sampled. Blood was
sampled from the caudal vein of the
individual fish after anaesthetization. The
whole blood was collected in a syringe,
Novriadi et al., 2015, Effect of Diets Containing Fish Protein 2

allowed to clot for 1 h in microtubes at room
temperature and followed by 5 h at 4 °C,
and then serum was harvested by
centrifuging at 1500×g for 5 min at 4 °C. All
serum samples were preserved at −20 °C
prior to analysis. The number and
percentage of leukocyte count, neutrophile,
monocyte and phagocytic was determined
based on Anderson and Sewicki (1993);
Blaxhall PC (1972) and Wedemeyer and
Yasutake (1977).
Water Quality Analysis
Ammonia and Posphate were
determined spectrophotometrically at 560
and 640 nm, respectively (Palkin Elmer©
Lambda XLS), Nitrite and Nitrate were
determined colorimetrically (HACH DR 890),
Turbidity, pH, dissolved oxygen, salinity and
temperature were determined by using
turbidimeter (Thermo Scientific), pH meter
(oakton), DO meter (oakton), refractometer
(ATAGO) and thermometer (Oakton),
respectively.
Total number of bacteria and Vibrio
Analysis
Every week, approximately 150 mL of
water from each rearing tank were sampled
to count bacteria, according to the methods
previously described (Gatesoupe, 1995).
Under sterile conditions, 1 mL of sample
was put in to three different dilutions, namely
103
, 102
and 101
. Dilution was performed by
using sterile Trisalt solution. After
homogenization, appropriate dilutions were
inoculated on Plate Count Agar (PCA, AES
Laboratoire, enriched with 18 g L-1
NaCl, pH
adjusted to 7.8) for total number of bacteria
analysis and TCBS agar (thiosulfate-citrate-
bile-salt agar, AES Laboratoire, dissolved in
half-strength seawater) for rough estimation
of bacteria and Vibrio spp. counts. The
plates were incubated for 24 hours at 28⁰ -
30⁰ C and counting was performed by using
colony counter.
Challenge test
Challenge test was performed by
using “bath challenge” with V.
parahaemolyticus at a density of 105
cells/mL after termination of the feeding trial
of the nursery. Twenty fish from each
supplemented and control group replicate
(±20 g) were immersed in V.
parahaemolyticus suspension for six hours.
Fish were then transferred to aquaria and
observed for five days for any clinical
abnormalities and mortalities.
Bacterial Culture
Isolates of the bacterial strains V.
parahaemolyticus obtained from
Brackishwater Research Centre, Jepara that
previously stored in 30 % glycerol at -80 0
C,
were aseptically inoculated in 30 mL marine
broth by incubation overnight at 25-28 0
C
with constant agitation. 150 µL was
subsequently transferred and grown to
stationery phase in 30 mL marine broth six
hours before challenge. The bacterial
densities were determined
spectrophotometrically at an optical density
of 550 nm. The bacterial densities were
calculated using the equation: Concentration
(CFU/mL) = [1200*106
*OD] according to
McFarland standard, assuming that an
OD550 = 1.000 corresponds to 1.2×109
cells/mL
Bacterial Stock
1 ml of the bacterial colony was
transferred and grown to stationery phase in
5 mL of Difco™ Marine Broth 2216 by
incubation overnight at 25-28 0
C with
constant agitation. Bacterial suspensions
were then transferred to centrifugation tubes
and centifugated at 4000 g for 5 minutes.
The supernatant was discarded and pellets
were resuspended in 7 mL filtered
autoclaved sea water (FASW). The solution
was homogenized and 3 ml of 30% Glycerol
solution was added. 150 µL of each colony
was distributed to the sterilized eppendorf
tube and stored at -80 0
C.
Statistical analysis
Data for growth and immune
performance of L. calcarifer are presented
as mean values followed by the standard
deviation. Survival data of L. calcarifer were
arcsine transformed for statistical
comparisons to satisfy normal distribution
and homoscedasticity requirements.
Survival data were subjected to one way
ANOVA followed by Tukey’s multiple
comparison range using the statistical
software SPSS version 21.0 to determine
significant differences among treatments. All
significance levels of the statistical analysis
were set at p<0.05.
3 Omni-Akuatika Vol. XIV No. 20 Mei 2015 : 1 - 12

RESULTS AND DISCUSSION
Growth Performance
In the current study, the influence of
functional hydrolysates in Asian seabass
during the nursery phase including total
weight gain (TWG), % relative weight gain
(% RWG), % specific growth rate (% SGR),
Final weight (g) and Final length are
summarized in Table 1.
Table 1. Growth performances of L. calcarifer fed dietary treatments for 6 weeks during the
nursery phase.
Treatments TWG (g) RWG (%) SGR (%) Final weight
(g)
Final length
(cm)
AQUATIV 3
% FPH
18.25±0.47
a
530.35±44.39
a
6.57±0.25
a
21.71±0.39
a
11.76±0.21
a
AQUATIV 2
% FPH
16.64±0.45
b
483.65±39.81
b
6.29±0.25
a
20.10±0.39
a
11.00±0.17
a
Control 9.87±0.89
c
287.13±35.99
c
4.82±0.34
b
13.33±0.84
b
10.33±0.32
b
TWG: Total Weight Gain (g), RWG: Relative Weight Gain (%), SGR: Specific Growth Rate
Significant differences among the treatments and control are indicated by different letter (n=2, P<0.05).
Following from this initial nursery phase, the dietary test went on for another 8 weeks in grow-out units.
Table 2. Growth performances of L. calcarifer fed dietary treatments for 9 weeks during the
grow-out phase.
Treatments TWG (g) RWG (%) SGR (%) Final weight (g) Final length
(cm)
AQUATIV 3
% FPH
40.74±1.23
a
187.80±7.40
a
1.92±0.05
a
62.94±1.47
a
16.56±0.23
a
AQUATIV 2
% FPH
36.76±2.60
b
182.99±13.84
a
1.89±0.09
a
56.94±2.26
b
15.55±0.28
b
Control 26.31±1.99
c
198.77±26.21
a
1.98±0.15
b
39.60±1.49
c
13.77±0.31
c
TWG: Total Weight Gain (g), RWG: Relative Weight Gain (%), SGR: Specific Growth Rate
During the grow-out feeding trial, SGR were lower for supplemented diet fish groups due to their higher
average weight and length. TWG however remained higher for fish groups receiving FPH supplemented
diets.
Immune Performance
Blood white cell counts are detailed in Table 3 (nursery) and Table 4 (grow out).
Table 3. Cellular immune response of L. calcarifer fed dietary treatments for 6 weeks during
the nursery phase.
Parameter Control
FPH Treatments
2 % 3 %
Neutrophile (%) 5.07±0.19
b
6.31±0.15
a
6.73±0.12
a
Monocyte (%) 1.93±0.2
b
2.65±0.1
a
2.8±0.13
a
Lymphocyte (%) 50.87±1.49
b
63.74±1.19
a
66.71±0.71
a
Macrophage (10
5
cell/mL) 1.83±0.25
b
2.62±0.12
a
2.83±0.15
a
Leukocyte (10
3
cell/mL) 48.33±1.36
b
55.67±1.14
a
56.34±1.17
a
Phagocytes activity (%) 15.77±1.18
b
37.95±1.01
a
40.23±0.52
a

Table 4. Cellular immune response of L. calcarifer fed dietary treatments for 9 weeks during
the grow-out phase.
Parameter Control
FPH Treatments
2 % 3 %
Neutrophile (%) 6.03±0.12 6.60±0.10 6.63±0.12
Monocyte (%) 2.40±0.10 2.87±0.12 2.93±0.06
Lymphocyte (%) 55.21±0.76 65.25±1.09 66.30±1.07
Macrophage (105
cell/ml) 2.30±0.20 2.60±0.10 2.67±0.12
Leukocyte (103
cell/ml) 49.68±0.56 57.01±2.36 54.67±0.43
Phagocytes activity (%) 28.29±0.67 39.64±1.03 40.48±0.95
Survival Rates
As illustrated by Figure 1, at the end
of nursery feeding trial, the 2 and 3%
hydrolysate supplementations resulted in
96.75±0.28 % and 97.28±0.18 % survival
rate respectively, while the control diet
yielded 93.65±0.13 %. More contrasted
results were observed at the end of the
grow-out feeding trial, most likely resulting
from a disease outbreak, which impacted
more the control diet group (20.1±21.1%)
compared to the supplemented diet groups
(78.4±7.7% and 86.0±4.32 % for 2 and 3%
FPH supplementation respectively).
Figure 1. Histogram of the mean percentage survival (%) of L. calcarifer during experiment
period in nursery phase (a) and grow out phase (b). Significant differences among the
treatments and control are indicated by different letter (p< 0.05).
A
B

Resistance to the bacterial challenge
At the end of the nursery feeding trial,
fish from each experimental groups (±20 g,
n=20) were challenged by immersion with V.
parahaemolyticus at a density of 105
cells/ml
and survival was observed for 5 days.
Figure 2 indicated that the supplementation
of functional hydrolysates was able to
induce significantly (p<0.05) higher survival
rate in comparison to control diet. In
addition, no significant difference was
observed between 2 % and 3 % application
of functional hydrolysates (p<0.05) even
though the 3% FPH supplemented group
yielded the highest survival rate with
78.33±2.89 %.
Figure 2. Histogram of the mean survival (%) of L. calcarifer (n=20, weight: ±20g) challenged
with V. parahaemolyticus at 105
cells/ml. Survival was scored after 5 days challenge with V.
parahaemolyticus. Significant differences among the treatments and control are indicated by
different letter (n=2, p< 0.05).
Water Quality Analysis
Table 5 Water quality analysis during AQUATIV nursery feeding trial. Sampling campaign was
performed on weekly basis at 9 AM at the middle point of experimental cages.
Paramet
er
Unit
Test Results
Method
SpecificationControl 2 % 3 %
Total
Bacteria
CFU/m
L
1.1 – 2.7x102
1.4 – 2.7x102 1.1 –
2.8x102
IKM/5.4.10/BBL-
B
Total
Vibrio
CFU/m
L
0.2 – 1.1x102
1.4 – 2.7x102 1.1 –
2.8x102 Convensional
pH 7.76 – 8.25 7.72 – 8.18 7.73 – 8.19
SNI 06-6989.11-
2004
Nitrate
(NO3)
mg/L <0.01
<0.01 <0.01
Colorimetry
Nitrite
(NO2)
mg/L <0.1
<0.1 <0.1
Colorimetry
Ammonia
(NH3)
mg/L <0.02 –0.094
<0.02 –0.094 <0.02 –0.094
IKM/5.4.6/BBL-B
Posphate
(PO4)
mg/L 0.012–0.024
0.018-0.027 0.015-0.022
IKM/5.4.8/BBL-B
Salinity ‰ 30 – 31 30 – 31 30 - 31 IKM/5.4.4/BBL-B
Turbidity NTU 0.65 – 1.42
0.81 – 1.44
0.65 – 1.42
IKM/5.4.9/BBL-B
Temperat
ure
⁰C 30.1 – 30.4
30.1 – 30.4 30.1 – 30.4
Thermometer

Table 6. Water quality analysis during AQUATIV grow-out feeding trial. Sampling campaign
was performed on weekly basis at 9 AM at the middle point of experimental cages.
Parameter Unit
Test Results
Metoda
Analisa
15 Juli 22 Jul 29 Jul 5 Aug 12 Aug 19
Aug
26 Aug
Total
Bacteria
CFU/
mL
173 1.2x10
3
5.2x10
3
1.4x1
0
3
2x10
2
2.7x1
0
2
3.7x10
3
IKM/5.4.10
/BBL-B
Total
Vibrio
CFU/
mL
23 131 2.7x10
2
3.4x1
0
2
1.9x10
2
3.1x1
0
2
1.9x10
2
Convensio
nal
pH
8.18 7.78 8.26 7.76 8.18 8.37 8.25 SNI 06-
6989.11-
2004
Nitrate
(NO3)
mg/L
<0.01 <0.01 <0.01 <0.01 <0.01 <0.01 <0.01 Colorimetr
y
Nitrite
(NO2)
mg/L
<0.1 <0.1 <0.1 <0.1 <0.1 <0.1 <0.1 Colorimetr
y
Ammonia
(NH3)
mg/L
<0.009 <0.009 <0.009 0.357 <0.009 <0.00
9
<0.009 IKM/5.4.6/
BBL-B
Posphate
(PO4)
mg/L
0.027 0.025 0.047 <0.03
3
<0.033 <0.03
3
<0.033 IKM/5.4.8/
BBL-B
Salinity ‰
30 30 31 30 30 31 31 IKM/5.4.4/
BBL-B
Turbidity NTU
7.15 2.63 5.33 27 2.44 6.52 7.26 IKM/5.4.9/
BBL-B
Temperatu
-re
⁰C
29.1 30.5 28.7 29.6 30.5 29.7 30.7 Thermom
eter
While observed water quality was
controlled and satisfying during nursery
feeding trial (Table 5), it was not possible to
guarantee a good water quality during the
grow-out feeding trial (Table 6). Water
quality observed during this latter period is
however representative of conditions which
may apply to any Asian seabass cage
rearing within this area. Most impacted
water quality indicators were the water
turbidity, i.e. the load of organic matters and
their associated pollutants and bacterial
communities, including a high load of Vibrio
spp. As illustrated above, degraded water
quality may be the origin of the disease
outbreak, which occurred during grow-out
feeding trial.
DISCUSSION
Protein hydrolysis is one promising
approach to improve the physiochemical,
functional, sensory and nutritional properties
of marine by product native proteins (Šližytė
et al., 2005). Many studies have revealed
that protein hydrolysis can also improve
intestinal absorption (Kristinsson and Rasco,
2000a) and microbiota (Kotzamanis et al.,
2007). Furthermore, hydrolysates are more
readily digested and absorbed in fish
digestive tract (Carvalho et al., 1997). In
order to replace fish meal, FPH are also
widely used as growth enhancers,
attractants or palatability enhancer (Hardy,
1991; Aguila et al., 2007). Several authors
have reported that good functional
properties and nutritive value of protein
hydrolysates improve the growth and feed
utilization in salmonids (Berge and
Storebakken, 1996; Refstie et al., 2004) and
in carp larvae (Carvalho et al., 1997). In line
with our results, at nursery phase, significant
increase of total weight gain (TWG), %
relative weight gain (% RWG), % specific
growth rate (% SGR), final weight (g) and
final length (cm) were recorded in Asian
Seabass treated with 2 % and 3 % of fish
hydrolysates in comparison to control
(p<0.05). The positive effects of functional
hydrolysates on Asian seabass
performances at nursery phase might be
explained by the high palatability of FPH
stimulating the feed intake (Cahu et al.,
1999; Oliva- Teles et al., 1999; Aguila et al.,
2007). It resulted in a higher growth rate with
higher biomass production. Based on the
significant differences observed for
zootechnical performances of Asian
Seabass at nursery phase (Table 1, 2 and
Figure 1), these functional properties of
hydrolysates obviously impacted the
nutritional metabolism of fish. During

hydrolysate manufacturing process, the
capability of enzymatic hydrolysis to
decrease the protein size of the raw material
and to improve the dietary protein quality
bring another advantage of using diet
containing functional hydrolysates
(Petersen, 1981). Protein hydrolysates will
be better absorbed than classical raw
materials such as meal (Cissé et al., 1995;
Ouellet et al., 1997), resulting in a higher
feed efficiency and less waste released into
the environment. Survival rate, relative
weight gain and specific growth rate in fish
fed functional hydrolysates were
considerably higher during the nursery
phase compared to the grow-out phase.
Under the controlled nursery conditions,
hydrolysates expressed their full potential. In
the grow out phase, fish were exposed to
persistent environmental challenges
(turbidity, temperature and salinity
fluctuations). Those adverse environmental
conditions actually resulted in a disease
outbreak and slowed growth down as
described in other studies (Walters and
Plumb, 1980; Robertson et al., 1987). The
condition developed by fish in the test
strongly relates to a Tenacibaculum infection
further complicated by opportunistic
vibriosis.
There have been reports of
biologically active peptides with immuno-
stimulating and antibacterial properties
being produced during the hydrolysate
manufacturing process (Coste et al., 1992;
Bøgwald et al., 1996; Gildberg et al., 1996;
Daoud et al., 2005; Kotzamanis et al., 2007).
The results of the present study showed that
feeding Asian Seabass with functional
hydrolysates enhanced phagocytosis activity
by blood phagocytic cells during the whole
experimental period and stimulated the
circulating neutrophile, monocyte,
lymphocyte and leukocyte number.
Observations on macrophages showed that
the application of 3 % of functional
hydrolysates was the most effective dose
among the treatments. It could increase the
number of macrophages after the natural
bacterial challenge. Macrophages play a
major role in both innate and adaptive
immune system. Study from Norum et al.
(2005) and Joerink et al. (2006) stated that
in the innate immune system, macrophages
act as phagocytes cells, produce pro
inflammation, ROS (by NADPH oxidase
enzyme) and RNS (by nitric oxide (NO-)
synthase). Meanwhile, in the adaptive
immune system, macrophages act as
professional APCs (Antigen presenting
cells). Moreover, the ability of macrophages
to kill pathogenic microbes is probably one
of the most important mechanisms of
protection in aquatic organisms (Selvaraj et
al., 2005). Corroboration for our results
comes from the work of Tang et al. (2008)
who have reported that growth performance
and immune parameters of the large yellow
croaker can be improved by supplementing
functional hydrolysates to their basal diet.
CONCLUSION
In conclusion, this work revealed that
the addition of functional hydrolysates in
Asian Seabass feed was able to enhance
significantly the fish immune system as well
as the fish zootechnical performances such
as Survival rate, total weight gain (TWG),
relative weight gain (RWG) and specific
growth rate (SGR) during the experimental
period. Therefore, dietary supplementation
of functional hydrolysates in Asian Seabass,
Lates calcarifer feeds should be encouraged
as a holistic approach to improving its
sustainable culture.
ACKNOWLEDGMENTS
This study was supported by SPF
DIANA (Member of SYMRISE Group),
through its AQUATIV Division by providing
operational fund, commercial feed and
functional hydrolysates to conduct field test
in Batam Mariculture Development Center.
We are thankful to the Reference laboratory
of Fish Diseases and Environmental
Analysis Serang for their assistance in
carrying out the histology analysis and
BMDC Laboratory staff for their assistance
in carrying out the water quality and
bacteriology analysis.
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